Hemoglobin Solution (1 mg/mL) - BioReagent

Cat. No.: H1528788

GRADE & PURITY BioReagent

Storage

Store at 2-8°C

Shipped In

Wet ice

Overview

This product is designed as a positive control for the qualitative detection of fecal occult blood. The formulation consists primarily of bovine hemoglobin, deionized water, and a preservative system. This reagent is supplied for research purposes exclusively and is not indicated for clinical diagnostic procedures or any other applications.

Specifications

Specifications & Purity

BioReagent

Storage

Store at 2-8°C

Shipped In

Wet ice

This product requires cold chain shipping. Ground and other economy services are not available.

Grade

BioReagent

Product Line

biological_products

Application Protocols

• Item-specific validated applications: Not specified for this item; refer to CoA/Spec Sheet.
• Example protocols (literature/general; adapt to your system)
  1. UV–Vis characterization of hemoglobin state
  • Dilute the 1 mg/mL stock to 0.05–0.2 mg/mL in air-equilibrated phosphate buffer (pH 7.4).
  • Record spectra 350–650 nm. OxyHb typically shows Soret ~413–418 nm and α/β bands ~576/540 nm. Confirm absence of turbidity by checking baseline scatter.
  2. Preparation of deoxyhemoglobin
  • Sparge buffer and sample with N2/Ar for 10–15 min on ice.
  • Add freshly prepared sodium dithionite (final 0.1–1 mM). Mix gently and cap; verify spectral shifts (Soret position and reduced Q-bands).
  3. Cyanomethemoglobin (Drabkin-type) reference (for teaching)
  • Combine an aliquot with Drabkin’s reagent per literature instructions; incubate until color stabilizes; read at 540 nm against reagent blank. Use for qualitative demonstration only unless calibrated with appropriate standards.
  • Note: Exact extinction coefficients and calibration factors vary by species and preparation; determine empirically for quantitative work.

Biological Roles

• General biochemistry (literature; no clinical claims)
  • Hemoglobin is a tetrameric oxygen-transport protein in vertebrate erythrocytes. Each subunit binds one O2 via an Fe(II)-heme, enabling cooperative ligand binding and efficient O2 delivery.
  • Allosteric regulation: Ligand binding shifts equilibrium between T (tense) and R (relaxed) states; proton concentration (Bohr effect), CO2, and organic phosphates (e.g., 2,3-BPG in humans) modulate affinity.
  • Redox states: Functional ferrous Hb(Fe2+) binds O2; oxidation generates methemoglobin Hb(Fe3+), which does not bind O2 but has distinct spectroscopic features; reduction systems (enzymatic in vivo) restore Fe(II).
  • Ligand specificity: Hemoglobin binds diatomic gases (O2, CO, NO) with differing affinities; competitive binding enables instructive kinetic and thermodynamic studies.
  • Structural biology: The globin fold is predominantly α-helical with a conserved heme pocket; numerous crystal structures provide models for protein–ligand interactions.
• Relevance of a 1 mg/mL solution
  • At this concentration, hemoglobin is well within the range for UV–Vis characterization, kinetic assays, and method development without excessive inner-filter effects (pathlength dependent).

Buffer Applications

• Not typically applicable: Hemoglobin is not used as a buffering agent.
• Practical note (literature)
  • Hemoglobin solutions are commonly prepared in neutral buffers (e.g., 50–100 mM phosphate, HEPES, or Tris, pH 7.0–8.0) to maintain structural integrity and defined ligand states; however, the exact buffer for this item is Not specified for this item; refer to CoA/Spec Sheet.

Green Alternatives

• Context: As an aqueous protein solution, this product already employs water as the primary solvent, aligning with green chemistry preferences for benign media.
• Comparative considerations (general)
  • For heme redox or catalysis studies where organic solvents are traditionally used, consider water-based or buffer-based systems with surfactants or co-solvents at minimal percentages.
  • If hemoglobin is used purely as a heme source, greener small-molecule alternatives (e.g., hemin chloride in aqueous buffers with minimal DMSO) can reduce protein waste and improve recyclability.
• Trade-offs
  • Hemoglobin provides biologically relevant cooperativity and allostery not replicated by simple heme models. Conversely, small-molecule heme analogs offer higher stability and solvent flexibility, reducing cold-chain energy demands.
• Summary table (literature/general)
  • Hemoglobin solution: pros — water-based, low VOC; cons — cold storage, limited lifetime, sensitive to oxidation.
  • Hemin/protoporphyrin: pros — longer shelf life, scalable in aqueous/green co-solvents; cons — lacks protein context and allosteric behavior.

Pharmaceutical Uses

• Not applicable to this catalog item.
• Context (general, non-clinical)
  • This BioReagent-grade hemoglobin solution is provided for laboratory research only. It is not formulated or qualified as an excipient or active ingredient. No pharmacopeial status is claimed. Do not use for diagnostic, therapeutic, or clinical purposes.

Physical Properties

• Item-specific (from Product Data)
  • Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Solution matrix: Not specified for this item; refer to CoA/Spec Sheet.
  • Exact pH, ionic strength, conductivity: Not specified for this item; refer to CoA/Spec Sheet.
• Literature/general values (protein context; not item specifications)
  • Hemoglobin (human HbA) molar mass: ~64.5 kDa (tetramer), each heme ~616 Da (literature).
  • Solubility: Highly soluble in aqueous buffers; insoluble in most organic solvents (literature).
  • Isoelectric point (pI): ~6.8–7.0 depending on species and buffer (literature).
  • Spectral properties (UV–Vis, heme Soret band): strong Soret band ~413–418 nm; Q-bands ~540 and ~576 nm for oxyHb; metHb ~405 nm; deoxyHb Soret slightly red-shifted (literature).
  • Extinction coefficients (illustrative, literature; vary by species/state): ε415 (oxyHb) ~1.2–1.3 × 10^5 M−1·cm−1 per heme; mass-specific A415 ~0.8–1.1 (mg/mL)−1·cm−1 for typical Hb preparations.
  • Stability: Sensitive to oxidation (Fe(II) → Fe(III), methemoglobin) and to denaturation at elevated temperatures or extreme pH (literature).

Quality & Grades

• Item-specific
  • Grade: BioReagent — suitable for general biological research applications where protein integrity and low bioburden/contaminants are important. Exact acceptance criteria (e.g., purity, endotoxin/bioburden limits, buffer composition, stabilizers) are Not specified for this item; refer to CoA/Spec Sheet.
  • Concentration: 1 mg/mL nominal. Lot-specific concentration and method of quantification (A415/A540, BCA, etc.) Not specified for this item; refer to CoA/Spec Sheet.
  • Storage: 2–8°C; shipped on wet ice (per Product Data).
• Practical implications of BioReagent grade (general)
  • Intended for research use in biochemical assays, spectroscopy training, oxygen-binding studies, and enzymology of heme proteins.
  • Typically filtered and prepared in defined buffers to reduce particulates; however, filtration status, preservative content, and stabilizers are Not specified for this item; refer to CoA/Spec Sheet.
  • UV background and aggregate content can influence spectroscopy; check lot-specific absorbance profile and turbidity before quantitative use.

Reaction & Applications

• Item-specific
  • Manufacturer applications: Not specified for this item in the catalog; see below for common research uses of hemoglobin solutions.
• Literature/general research applications
  • Spectroscopic standards: Strong Soret/Q-band absorbance makes hemoglobin a convenient calibrant and teaching standard for UV–Vis spectroscopy. Control of O2/CO/NO binding states enables distinct spectral signatures.
  • Oxygen-binding/allostery studies: Evaluate cooperative binding (Hill plots), Bohr effect (pH dependence), and effects of salts or small molecules (e.g., 2,3-BPG in human Hb).
  • Enzymology and redox chemistry: Peroxidase-like activity of heme permits kinetic assays with H2O2 and chromogenic donors; study autoxidation to methemoglobin.
  • Gas-binding kinetics: Stopped-flow or rapid-mixing experiments with CO/NO/O2 for association/dissociation rate determination.
  • Analytical chemistry: Model matrix for hemolysate analysis, method development for heme quantification, or as a colored protein control in fractionation workflows.
  • Materials/biomimetics: Incorporation into hydrogels/membranes for gas-sensing prototypes (note: verify compatibility and stability empirically).

Reaction Conditions

• Item-specific: No standardized reaction conditions are provided for this item; refer to CoA/Spec Sheet for any lot-specific guidance.
• General experimental conditions for hemoglobin studies (literature; examples)
  • Oxygenation/deoxygenation: Buffer pH 7.2–7.6, 20–25°C; deoxygenate by sparging with N2/Ar and adding a stoichiometric reductant (e.g., sodium dithionite) to prepare deoxyHb; reoxygenate by gentle O2 bubbling.
  • CO/NO binding kinetics: Perform in airtight cuvettes or stopped-flow; maintain low light and controlled gas partial pressures; temperature 10–25°C. Monitor at 415, 430, 540, 576 nm (state-dependent maxima).
  • Peroxidase-like assays: 10–100 µg/mL Hb in phosphate buffer (pH 7.0–7.4) with H2O2 (10–500 µM) and chromogenic substrates (e.g., ABTS, guaiacol); track increase at 405–420 nm. Optimize to avoid heme degradation.
  • Redox interconversion: MetHb generation with mild oxidants (e.g., potassium ferricyanide), followed by removal (desalting) and spectral verification (Soret ~405 nm for metHb).

Safety & Handling

• Item-specific hazard information (from Product Data)
  • Signal Word: Not specified for this item; refer to SDS.
  • GHS Classification / Pictograms / H-Statements: Not specified for this item; refer to SDS.
• General laboratory safety (protein solutions; literature/general guidance)
  • Expected hazards: Hemoglobin solutions are generally considered low hazard for typical laboratory handling but may cause irritation upon contact. Avoid inhalation of aerosols and contact with skin/eyes.
  • PPE: Use lab coat, gloves (nitrile), and safety glasses. Handle in a clean environment to maintain sample integrity.
  • Incompatibilities: Strong oxidizers (promote methemoglobin formation), strong acids/bases (protein denaturation), and metal chelators that may perturb heme coordination.
  • First aid (overview; defer to SDS): Rinse skin/eyes with water if contacted. If inhaled as aerosol, move to fresh air. Seek medical attention if symptoms persist.
  • Spill/cleanup: Absorb with inert material, clean with aqueous detergent; decontaminate surfaces with standard laboratory disinfectants.
  • Waste: Dispose according to institutional guidelines for nonhazardous biological/chemical aqueous waste.
  • Always consult the product’s SDS for authoritative safety data and institutional EHS requirements.

Solvent Selection

• Relevance: This product is an aqueous protein solution; organic solvent selection is generally not applicable.
• General guidance (literature)
  • Preferred media: Neutral aqueous buffers (e.g., phosphate, HEPES, Tris) at ionic strength ~50–200 mM help maintain hemoglobin solubility and native conformation. Avoid extremes of pH (<6 or >9) that can shift heme state or denature globin.
  • Additives: Gentle reducing systems (e.g., dithionite for deoxyHb preparation) or CO/O2 equilibration gases are used to control redox/ligand state. Chelators and strong oxidants disrupt function.
  • Temperature: Work at 2–8°C for stability; room temperature for short measurements is typical.
• When to choose alternatives
  • For nonaqueous or denaturing applications, use model heme compounds (e.g., hemin, protoporphyrin IX) rather than hemoglobin, as proteins are unstable in organic solvents.

Storage & Reconstitution

• Item-specific
  • Storage condition: Store at 2–8°C (per Product Data).
  • Shipping: Shipped on wet ice (per Product Data).
  • Form: Ready-to-use aqueous solution at 1 mg/mL. No reconstitution required.
  • Stabilizers/preservatives/buffer composition: Not specified for this item; refer to CoA/Spec Sheet.
• General handling guidance (literature; not item specifications)
  • Minimize freeze–thaw and repeated warming; keep on ice during use and return to 2–8°C promptly.
  • Work under low-light conditions to limit photochemical oxidation; consider aliquoting to reduce air exposure if repeated use is expected.
  • Monitor integrity by UV–Vis profile; formation of methemoglobin or precipitation indicates degradation.
  • For any storage beyond the labeled recommendations or for freezing, establish conditions empirically; consult the SDS/CoA for lot-specific advice.

Structure & Identity

• Item-specific (from Product Data)
  • Product: Hemoglobin Solution (1 mg/mL)
  • Grade: BioReagent
  • Concentration: 1 mg/mL (aqueous solution; exact buffer composition Not specified for this item; refer to CoA/Spec Sheet.)
  • CAS: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular Formula: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular Weight: Not specified for this item; refer to CoA/Spec Sheet.
  • SMILES / InChI / InChIKey: Not applicable for macromolecular protein mixtures; Not specified for this item; refer to CoA/Spec Sheet.
• Literature/general description (for context)
  • Hemoglobin is a tetrameric globular protein typically composed of two α and two β subunits (HbA in humans), each containing a noncovalently bound heme b prosthetic group with central Fe(II).
  • Overall quaternary structure shows cooperative O2 binding and allosteric regulation (T and R states).
  • Typical subunit sizes: α ≈ 141 aa; β ≈ 146 aa; total tetramer ≈ 64.5 kDa (human HbA, literature).
  • 2D structural features: porphyrin macrocycle (heme) is planar with conjugated pyrrole rings; protein domains are predominantly α-helical (globin fold).

Synthetic Utility

• Not a synthetic building block: Hemoglobin is a macromolecular protein solution and is not typically used as a reagent for bond-forming organic synthesis.
• Useful roles in chemical research (literature)
  • Heme-mediated catalysis model: The embedded heme can catalyze peroxidative transformations, serving as a biomimetic platform to study radical mechanisms and selective oxidations under aqueous conditions.
  • Analytical standard: Colored protein for calibrating spectrophotometers (visible region) and validating extraction/fractionation protocols in complex matrices.
  • Ligand-binding thermodynamics: Benchmark system for evaluating models of cooperativity and for testing data-analysis workflows (Hill plots, MWC models).

Target Specificity

• Not applicable: This product is not an antibody or affinity reagent. No target, clone, isotype, or species reactivity information applies.